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In this paper we demonstrate the use of recombinant viral vectors derived from herpes simplex virus type 1 (HSV1) to transfer reporter genes in vitro into rat anterior pituitary cells grown in primary cultures and the anterior pituitary tumour cell lines GH3 and AtT20. The three vectors used were, tsK/beta-galactosidase (beta-gal), tsK/CRH and tsK/TIMP, the corresponding transgene products respectively being E. coli beta-gal, pre-procorticotropin releasing hormone (ppCRH), and the chimeric protein TIMP/Thy1 (tissue inhibitor of metalloproteinases (TIMP)/linked to the carboxy terminus of Thy1 which confers the addition of a glycolipid glycosyl-phosphatidylinositol anchor in the ER). Double labelling immunofluorescence experiments to detect reporter proteins and transduced cell types indicated that the three vectors could transfer and express the reporter genes in normal and tumour anterior pituitary cells. Virus infection of pituitary cells was characterised, and it was shown that infection with tsK/beta-gal at multiplicities of infection (MOI)=10, 100% of tumour and non-endocrine anterior pituitary cells expressed beta-gal, whereas 75% endocrine anterior pituitary cells expressed the transgene. Long-term expression studies after infection with tsK/beta-gal indicated that anterior pituitary cells in primary cultures expressed the transgene for significant longer periods than tumour anterior pituitary cells. Growth arrest by serum starvation markedly decreased the frequency of transgene expression in anterior pituitary cells following infection with tsK/beta-gal. Transgenic products expressed from tsK were targeted to their correct intracellular domain in both anterior pituitary cells in primary cultures and in pituitary tumour cell lines. We conclude that transgenes can be delivered into anterior pituitary cells in primary culture and pituitary tumour cell lines using tsK derived HSV1 vectors. The prospect of employing viral vectors to transfer genes into endocrine cells opens up the potential exploration of various molecular aspects of pituitary cell function both in vitro and in vivo, as well as the use of gene transfer into the pituitary for potentially therapeutic applications, such as the treatment of pituitary tumours.
R G Goya, and
J Rowe, and
Y E Sosa, and
P Tomasec, and
P R Lowenstein, and
M G Castro
January 2015,
Methods in molecular biology (Clifton, N.J.),
R G Goya, and
J Rowe, and
Y E Sosa, and
P Tomasec, and
P R Lowenstein, and
M G Castro
January 1997,
Advances in neurology,
R G Goya, and
J Rowe, and
Y E Sosa, and
P Tomasec, and
P R Lowenstein, and
M G Castro
October 1996,
Journal of virology,
R G Goya, and
J Rowe, and
Y E Sosa, and
P Tomasec, and
P R Lowenstein, and
M G Castro
January 2015,
Methods in molecular biology (Clifton, N.J.),
R G Goya, and
J Rowe, and
Y E Sosa, and
P Tomasec, and
P R Lowenstein, and
M G Castro
January 2011,
Methods in molecular biology (Clifton, N.J.),
R G Goya, and
J Rowe, and
Y E Sosa, and
P Tomasec, and
P R Lowenstein, and
M G Castro
January 2000,
Folia morphologica,
R G Goya, and
J Rowe, and
Y E Sosa, and
P Tomasec, and
P R Lowenstein, and
M G Castro
April 2000,
Nihon rinsho. Japanese journal of clinical medicine,
R G Goya, and
J Rowe, and
Y E Sosa, and
P Tomasec, and
P R Lowenstein, and
M G Castro
January 1994,
Developments in biological standardization,
R G Goya, and
J Rowe, and
Y E Sosa, and
P Tomasec, and
P R Lowenstein, and
M G Castro
March 1997,
Nature medicine,
R G Goya, and
J Rowe, and
Y E Sosa, and
P Tomasec, and
P R Lowenstein, and
M G Castro
July 1999,
Human gene therapy,
